Global Patent Index - EP 0548056 B2

EP 0548056 B2 20010404 - A process for cloning methylated DNA in a bacterial host incapable of host restriction

Title (en)

A process for cloning methylated DNA in a bacterial host incapable of host restriction

Title (de)

Klonierung von methylierter DNS in einem bakteriellen Wirtsstamm der zur Wirtsrestriktion unfähig ist

Title (fr)

Processus pour le clonage d'ADN méthylée dans une souche bactérienne hôte qui est incapable de restriction

Publication

EP 0548056 B2 20010404 (EN)

Application

EP 93200367 A 19890718

Priority

  • EP 89201895 A 19890718
  • NL 8801826 A 19880719

Abstract (en)

[origin: EP0353812A2] A process for the rescue and cloning of a DNA sequence wherein a vector containing said DNA sequence is multiplied in a suitable bacterial host. As the bacterial host, a bacterial strain which is incapable of host restriction is used, such as an Escherichia coli C strain. Preferably, a bacteriophage vector which is a suitable cloning vehicle for the bacterial host concerned is used as the vector, the step of in vitro packaging into phage coats being performed by means of a phage packaging extract obtained from a bacterial strain which is incapable of host restriction. The above may be used in a process for detecting mutations in one or more marker genes introduced into a mammalian genome by means of a vector, which process comprises isolating DNA from cells of the transgenic mammal or from the mammalian cells, recovering the vector from said DNA, multiplying the vector in a suitable bacterial host which is deficient with respect to at least one of the marker genes, and testing marker gene expression. Preferably, the DNA isolated from cells of the transgenic mammal or the mammalian cells is pre-purified by fragmenting the DNA by means of a restriction enzyme which does not have a cutting site within the vector, separating the resulting fragments on the basis of a suitable criterion, such as differences in size, and collecting fragments comprising the vector, whereafter the vector is recovered from said pre-purified DNA.

IPC 1-7

C12N 15/00; C12N 15/10; C12N 15/73; C12N 15/85; C12Q 1/68

IPC 8 full level

A01K 67/027 (2006.01); C12N 1/21 (2006.01); C12N 15/09 (2006.01); C12N 15/10 (2006.01); C12N 15/73 (2006.01); C12N 15/85 (2006.01); C12Q 1/68 (2018.01); C12Q 1/6827 (2018.01); C12R 1/19 (2006.01)

CPC (source: EP)

C12N 15/10 (2013.01); C12N 15/73 (2013.01); C12N 15/85 (2013.01); C12Q 1/68 (2013.01); C12Q 1/6827 (2013.01)

Citation (opposition)

Opponent :

  • Raleigh. Restriktion and modification in vivo by Escherichia coli K12. Methods in Enzymology 12, 130-141 (1987)
  • Rosenberg et al. Improved in vitro packaging of coliphage lambda DNA; a one strain system free from endogenous phage. Gene 38, 165-175 (1985)
  • Rosenberg. EcoK restriction cloning during in vitro packaging of coliphage Lambda DNA. Gene 39, 313-315 (1985)

Designated contracting state (EPC)

AT BE CH DE ES FR GB GR IT LI LU NL SE

DOCDB simple family (publication)

EP 0353812 A2 19900207; EP 0353812 A3 19900307; EP 0353812 B1 19940112; AT 100151 T 19940115; AT 132529 T 19960115; DE 68912228 D1 19940224; DE 68912228 T2 19940804; DE 68925369 D1 19960215; DE 68925369 T2 19960515; DE 68925369 T3 20010809; EP 0548056 A1 19930623; EP 0548056 B1 19960103; EP 0548056 B2 20010404; EP 0683238 A2 19951122; EP 0683238 A3 19951220; ES 2061943 T3 19941216; ES 2081679 T3 19960316; GR 3018927 T3 19960531; JP H02142471 A 19900531; JP H0829093 B2 19960327; NL 8801826 A 19900216; US 5470706 A 19951128

DOCDB simple family (application)

EP 89201895 A 19890718; AT 89201895 T 19890718; AT 93200367 T 19890718; DE 68912228 T 19890718; DE 68925369 T 19890718; EP 93200367 A 19890718; EP 95201753 A 19890718; ES 89201895 T 19890718; ES 93200367 T 19890718; GR 960400333 T 19960207; JP 18719389 A 19890718; NL 8801826 A 19880719; US 1319893 A 19930129